1181
J. Med. Chem. 1985,28, 1181-1187
Analogues of Platelet Activating Factor (PAF). 2.' Some Modifications of the Glycerine Backbone A. Wissner,* R. E. Schaub, P.-E. Sum, C. A. Kohler, and B. M. Goldstein Metabolic Disease Research a n d Cardiovascular Biological Research Sections, Medical Research Division, American Cyanamid Company, Lederle Laboratories, Pearl River, New York 10965. Received December 7, 1984 Racemic analogues of platelet activating factor (PAF) that contain a methylene group between the C2and C3 carbon atoms (39) or between the C1 and C2 carbon atoms (40) have been synthesized. These compounds show reduced platelet aggregation and hypotensive activity as measured against racemic c16 PAF. Compounds in which the C1 carbon atom of PAF is substituted with one or two methyl groups (41 and 42, respectively) or the C3 carbon is substituted with a single methyl group (43) have been synthesized. Platelet aggregation and hypotensive responses produced by these compounds are significantly less than those obtained with racemic c16 PAF. None of the above compounds exhibit a separation of the platelet aggregation and hypotensive activities.
Platelet activating factor (PAF), an alkyl ether phospholipid comprised primarily of c16 and CIShomologues (la and lb), has a variety of interesting biological properties among which is its ability to activate various inflammatory cell types (e.g., platelets, neutrophils, and basophils) and to lower blood pressure.2
Scheme I CH3(CH2),,0H CICH2C02~
toluene,
A
-
%-
CH~(CHZ)~SOCHZCOZH
2 C 0 CI -Me SO
CH3(CH2)i~OCHzCHzOH &2&:
-2:.c
3
CH3(CHz),,0CH2CH0
CH$CH2)150CH2~HCH=CHz
I
9
4 OH
CH3CO 2$2.i
e
II
C6H5CHzBr, DMF
NOH
-
5
3 CH2-OPOCHzCHzN(CH3)3'
I
0-
la, n.15 b. n.17
Previous research on PAF analogues have involved the preparation of compounds in which the length of the alkyl chain was varied3 or substituted with an aromatic ring.l We h a v e reported a deoxy analogue in which the ether oxygen has been removed,' while others have described a compound in which a sulfur atom replaces the ether oxygen.4 Compounds have been prepared in which the 2acetyl g r o ~ p o~r ,the ~ p h o s p h ~ c h o l i n eportion ~ ~ ~ of the molecule has been modified. In a continuation of our effort to prepare a more selective analogue that maintains good antihypertensive activity and has diminished potency as a platelet aggregating agent, we have prepared a number (1) For previous publication in this series, see: Wissner, A.; Sum,
P.-E.; Schaub, R. E.; Kohler, C. A.; Goldstein, B. M. J . Med. Chem. 1984,27, 1174. (2) For recent review of the biological properties of PAF see: (a) Snyder, F. Annu. Rep. Med. Chem. 1982,17,243. (b) Pinckard, R. N.; McManus, L. M.; Hanahan, J. Adu. Inflammation Res. 1982,4, 147. (c) Vargaftig, B. B.; Chignard, M.; Benveniste, J.; Lefort, J.; Wal, F. Ann. N.Y. Acad. Sei. 1981,370, 119. (3) (a) Hanahan, D. J.; Munder, P. G.; Sartouchi, K.; McManus, L.; Pinckard, R. N. Biochem. Biophys. Res. Commun. 1981,99, 183. (b) Tence, M.; Coeffier, E.; Heymans, F.; Polonsky, J.; Godfroid, J.; Benveniste, J. Biochim. 1981, 63, 723. (4) Hillmar, I.; Muramatsu, T.; Zollner, N. 2.Phys. Chem. 1984, 365, 33. ( 5 ) (a) O'Flaherty, J. T.; Salzer, W. L.; Cousort, S.; McCall, C. E.; Piantadosi, C.; Suries, J. R.; Hammett, M. J.; Wykle, R. L. Res. Commun. Chem. Pathol. Pharmacol. 1983,39,291. (b) Blank, M. L.; Cress, E. A.; Lee, T.-C.; Malone, B.; Surles, J. R.; Piantadosi, C.; Hajdu, J.; Snyder, F. Res. Commun. Chem. Pathol. Pharmacol. 1982,38, 3. ( c ) Wykle, R. L.; Surles, J. R.; Piantadosi, C.; Salzer, W. L.; O'Flaherty, J. T. FEBS Lett. 1982, 141, 29. (d) Chandrakumar, N. S.; Hajdu, J. J. Org. Chem. 1982,48, 1197. (6) Sartouchi, K.; Pinckard, R. N.; McManus, L. M.; Hanahan, J. T. J.Biol. Chem. 1981, 256, 4425. (b) Moschidis, M. C.; Demopoulos, C. A.; Kritikou, L. G. Chem. Phys. Lipids 1983,33, 87.
0022-2623/85/1828-1181$01.50/0
" 7
Scheme I1 cp,H
r0(cH2)15C CO(CH2)15cH3 (C6H5)3CCI pyridine
(1) C&CH29r, NaH, OMF ( 2 ) PTSA, CtMH-THF
L O H 10
of analogues that incorporate alterations of the glycerine backbone of the molecule. Specifically, we report the syntheses and biological activities of racemic analogues in which the glycerine backbone has been substituted at the C1 or C3 carbon atoms with methyl groups and molecules in which the length of the backbone has been increased b y the addition of a methylene group between the Cl-C2 or C2-C3 bonds. Chemistry. The various synthetic strategies developed for the preparation of the PAF analogues all converge t o similar intermediates (7, 12, 23, 24, 27) that contain the alkyl ether chain, a secondary benzyloxy group, and a free 0 1985 American Chemical Society
1182 Journal of Medicinal Chemistry, 1985, Vol. 28, No. 9 Scheme I11
Wissner et a [ . Scheme IV
R
rO(CH2)15CH3 (COCI12-Me2S0
b0cH2c6H5
CHZC12, -IO 'C
0CH2C6H!3
ether-THF
-OH 13, R=H 14. R=CH3
25
A
R
R (11 CH,C%H,
O(CHz)15CH3
PTSA, A
-
(21 CHQOH, H20. NoOH or KOH. MepSO
15, R = H 16, R=CH3
17, R = H 18. R=CH, (21 NaOAc, H20
(3) (CH313N, CH3CN, CHCI3. A CC(C,H,13
or CC(C6H512C6H4-p pyridine
- OCH3
c
7, 12, 23, 24.
R,=R2=R3=H. 0.1. m.2 R,=R =R3=H. n.2, m.1 R1=Ch3, R 2 = R 3 = H . n = l , m = l R i = R 2 = C H j . R =H, n = l , m = l 27. Rl=R2=H. R3=?H3. n = l , m = l
LOH
19. R = H
20. R=CH3
Lox 21. R=H. X=C(C,H,), 22,R=CH3. X=C(C&),GH,-p-OCH,
23. R = H 2 4 , R=CH3
hydroxyl group. The syntheses of these precursors are detailed below. Introduction of a Methylene Group between the C2 and C3 Carbon Atoms (Scheme I). Alkylation of 1hexadecanol with chloroacetic acid is accomplished by the prior formation of the sodium salt of the acid by its addition to a suspension of sodium hydride in toluene; addition of the alcohol and refluxing for 40 h give the ca-boxylic acid 2. The aldehyde 4 is prepared by reduction of 2 with borane-THF to give alcohol 3 followed by oxidation of 3 with oxalyl chloride-Me2S0 at -20 "C. We have found 4 to be a rather unstable substance that is best stored a t low temperature in solution (petroleum ether, 30-60 "C) prior to use. The reaction of 4 with vinylmagnesium bromide gives the allylic alcohol 5. Protection of the hydroxyl group as a benzyl ether (6) followed by hydroboration-oxidation then gives the desired precursor 7.
Introduction of a Methylene Group between the C1 and C2Carbon Atoms (Scheme 11). Alkylation of 3buten-1-01 with 1-bromohexadecane using sodium hydride in DMF gives the ether 8. Oxidation of 8 with m-chloroperbenzoic acid furnishes the epoxide 9 that is then converted to diol 10 in a two-step process involving an acidcatalyzed epoxide ring opening in acetic acid followed by hydrolysis of the resulting mixture of monoacetates with methanolic sodium hydroxide. The primary hydroxyl group of 10 is selectively protected with trityl chloride in pyridine. Alkylation of the hydroxyl group of 11 with benzyl bromide and sodium hydride in DMF followed by removal of the trityl protecting group in an acidified mixture of MeOH and T H F then gives the desired precursor 12. Mono- and Dimethyl Substitution at the C, Carbon Atom (Scheme 111). Alkylation of 3-buten-2-01 or 2methyl-3-buten-2-01 with the tosylate of 1-hexadecanol using sodium hydride in dimethylformamide gives the allylic ethers 15 and 16, respectively. Both were oxidized
to the respective epoxides 17 and 18 with m-chloroperbenzoic acid in methylene chloride. Refluxing an acetic acid solution of 17 in the presence of of a catalytic amount of p-toluenesulfonic acid followed by basic hydrolysis of the resulting mixture of monoacetates gave the diol 19. On attempted acid-catalyzed ring opening of the more hindered epoxide 18, it was recovered unchanged. Successful ring opening was achieved however, using potassium hydroxide in MczSOat 110 "C for 8 h, giving the diol 20 along with a dimer side product (see Experimental Section). Diols 19 and 20 were then converted to the desired precursors 23 and 24, respectively, by the protection-alkylation-deprotection sequence described above. It was evident, by thin-layer chromatography, that both possible diastereoisomers of 23 are present in about equal amounts. Methyl Substitution at the C3 Carbon Atom (Scheme IV). Oxidation of 25* with oxalyl chlorideMe2S07in methylene chloride at -10 "C gave the aldehyde (7) Mancuso, A. J.; Huang, S.-L.; Swenn, D. J . Org. Chem. 1978. 43, 2480.
Analogues of Platelet Activating Factor
Journal of Medicinal Chemistry, 1985, Vol. 28, No. 9 1183 Table I. Blood Pressure and Platelet Aggregation Activities for
PAF Analogues
039, Rl=Rz=R3=H, n = l , m . 2 40. R1=R2=R3=H,n = 2 ,m = l 41, R,=CH3. R,=R3=H. n = l . m.1 42. R,=RZ=CH,. R3.H. n.1. m.1 43, Ri=RZ=H. R3zCH3, n=l. m.1
26. The reaction of 26 with methylmagnesium bromide in ether-tetrahydrofuran furnished the desired intermediate 27. Thin-layer chromatography indicated the presence of both possible diastereoisomers in about equal amounts. Preparation of PAF Analogues 39-43 (Scheme V). The phosphocholine groups were introduced with 2bromoethyl phosphorodichloridate (28) by a modification of the method of Hirt.g The reaction of a carbon tetrachloride solution of 7, 12, 23, 24, or 27 with an excess of 28 and triethylamine followed by hydrolysis in aqueous sodium acetate gave a series of bromoethyl phosphates that were converted to phosphocholines 29-33, respectively, by refluxing in a solution of chloroform-acetonitrile containing a large excess of anhydrous trimethylamine. The reaction of 28 with the more hindered alcohol 27 required a considerably longer reaction time (48 h) than the reaction with the less hindered alcohols. The benzyl protecting groups were removed from 29-33 by catalytic hydrogenolysis using 5% Pd/C in a mixture of acetic acid and MeOH. Finally, the alcohols 34-38 were acetylated with acetic anhydride and triethylamine in chloroform at reflux to give the PAF analogues 39-43 (Chart I). Biology. We have proposed the hypothesis that the various biological effects of PAF need not be mediated by the same receptor and that the hypotensive activity of PAF is not a consequence of mediator cell (e.g., platelet) activation and release.' If this is true and if the receptor responsible for cell activation and that responsible for the hypotensive effect differ sufficiently in their structural requirements, then it may be possible to prepare an analogue that retains the hypotensive activity yet has diminished activity with respect to cell activation and release. Such a compound may be a therapeutically useful hypotensive agent. Evidence that at least some of the biological activities of PAF are receptor-mediated processes include its stereospecificity of action,1° the discovery of a specific antagonist," and the actual identification of PAF receptors.12 To examine the effect of our PAF analogues on mediator cell activation, we have chosen to study platelet aggregation Arnold, D.; Weltzien, H. U.; Westphal, 0. Liebigs Ann. Chem. 1967, 709, 234. Hirt, R.; Berchtold, R. Pharm. Acta Helu. 1958, 33, 349. (a) Wykle, R. L.; Miller, C. H.; Lewis, J. C.; Schmitt, J. D.; Smith, J. A.; Surles, J. R.; Piantadosi, C.; O'Flaherty, J. T. Biochem. Biophys. Res. Commun. 1981,100,1651. (b) Tence, M.; Coeffies, E.; Polonsky, J.; Benveniste, J. Biochim. Biophys. Acta 1983, 735, 526. Terashita, Z.-i.; Tsushima, S.; Yoshioka, Y. Nomura, H.; Inada, Y.; Nishikawa, K. Life Sci. 1983, 32, 1975. (a) Valone, F. H.; Coles, E.; Reinhold, V. R.; Goetzl, E. J. J . Immunol. 1982, 129, 1637. (b) Valone, F. H.; Goetzl, E. J. J . Zmmunol. 1983, 48, 141. (c) Hwang, S.-B.; Lee, C.-S.; Cleah, M. J.; Shen, T. Y. Biochemistry 1983, 22, 4756.
max platelet EC6t aggregation compd (n) respc ratiod MABP" (7) 75 (1.3 X lov5) 0.65 la 1.25 (0.30, 4.98) 1.94 X 39 47.5 (10.6, 197.8) 1.4 X lo4 (1) 65 (5 X lo-,) 0.34 40 24.3 (9.3, 63.0) 1.5 X lo4 (1) 50 (1.9 X 0.16 41 7.12 (3.69, 13.9) 5.1 X (1) 65 (5 X 0.14 42 875.7 (229.2, 1.1 x 10-5 (1) 64 (5 x 10-5) 0.80 3564) 43 63.4 (13.2, 262.0) 3.2 X lo4 (1) 70 (1.9 X 0.20 "Dose (rg/kg, iv) required to decrease mean arterial blood pressure (MABP) 50 mmHg. Values in parentheses are 95% confidence limits. *Molar concentration required to produce 50% of maximum aggregation. The n values are the number of experiments in which a dose-response curve was determined from two to six replicates per dose level. Maximum aggregation units at the specified molar concentration. (MABP/platelet EC,,) x
using rabbit platelet-rich plasma. We have chosen the platelets of this species since they are known to be very sensitive to PAF.2cJ3 The data (Table I) are expressed as the molar concentration of the analogue required to obtain 50% of its maximum response (EC,,) and as the maximum aggregation response of the analogue obtained at the indicated molar concentration. This latter value is useful in order to distinguish partial from full agonists. Spontaneously hypertensive rats were used for blood pressure studies. It is known that the platelets of this species do not respond well to PAF and that the hypotensive effect is not mediated by platelets.2c Blood pressure data (Table I) are expressed as the intravenous dose of the analogue needed to reduce the mean arterial blood pressure (MABP) 50 mmHg as determined from a leastsquares regression line. The last column in Table I gives the ratio of the blood pressure and platelet aggregation values and can be used as a measure of the degree of separation of the two activities for a particular compound relative to the standard compound (racemic C16 PAF, la). A value of this ratio smaller than that observed for l a may indicate selectivity in favor of the hypotensive effect while a larger ratio would suggest selectivity in favor of platelet aggregation. Results and Discussion The blood pressure and platelet aggregation data obtained for racemic l a and analogues 39-43 are presented in Table I. Increasing the length of the glycerine backbone of PAF by the introduction of a methylene group between carbon atoms Cz and C3(compound 39) or between carbon atoms C1 and Cz (compound 40) results in a decrease of both the hypotensive (38- and 19-fold) and platelet aggregation (73- and 77-fold) responses, respectively, when compared to la. Methyl substitution a t both C1 (compound 41) and at C3 (compound 43) also result in a decreased response in both assays, and it is apparent that this effect is greatest for C3 substitution. It should be pointed out that these results are complicated by the fact that 41 and 43 are both a mixture of two diastereomers, with each diastereomeric component of the mixture containing an enantiomer with the same configuration as the natural isomer, and consequently it is not known to what extent the activities reside in one or both of the enantiomers of the mixture. Substitution of C1 with two methyl groups (compound 42) (13) (a) Benveniste, J.; Henson, P. M.; Cochrane, C. G. J . Exp. Med. 1972, 136, 1356. (b) Camussi, G.; Tetta, C.; Deregibus, M. C.; Bussolino, F.; Segoloni, G.; Vercellone, A. J . Zmmunol. 1982, 128, 86.
1184 Journal of Medicinal Chemistry, 1985, Vol. 28, No. 9
*
1 t
-
-
2
i.
1C
\
4
_r
c
3"
Figure 1. Relationship between the natural logarithms of the MABP and platelet ECSo values. Error bars are the natural logarithmic values of the 95% confidence limits for the MABP values. The regression equation is In [platelet EC,,] = 0 925 In [MABP] - 17.0
results in an even larger decrease in the hypotensive (700-fold) and platelet-aggregating (567-fold) activities. The ratios of the hypotensive and platelet-aggregating responses observed for 39-43 span a rather narrow range (0.14-0.80) and are comparable to that observed for the standard compound la (0.65),suggesting that these strucutral modifications have resulted in little or no significant changes in selectivity. These results are presented graphically in Figure 1in which the log values of the hypotensive and platelet-aggregating responses of la and 39-43 are plotted. A linear correlation is observed with an R2 value of 0.89. While we have not observed the hoped for hypotensive selectivity in this series of compounds, we are, nevertheless, continuing with our efforts in this area. Experimental Section General Methods. Unless otherwise stated, the following are implied. Melting points were determined on a Mel-Temp capillary melting point apparatus and are uncorrected. The nuclear magnetic resonance (NMR) spectra were recorded on either a Varian EM 390 spectrometer or Varian FT-80 spectrometer, and chemical shifts in parts per million (ppm) are reported with tetramethylsilane (Me4Si)or chloroform as internal references. Infrared spectra (IR) were recorded on a Nicolet F T 7000 spectrophotometer. Mass spectra were determined on a FinneganMAT Model CH 7 mass spectrometer. The field desorption (FD) (FD p-TSA), and fast atom bombardment (FAB) mass spectra were obtained on a Kratos MS 50 mass spectrometer. Where analyses are indicated only by symbols of the elements, analytical results obtained for these elements were within f0.4% of the theoretical value. Biological Assays. The methods used in the platelet aggregation and blood pressure assays have been described previously 1-(Hexadecy1oxy)acetic Acid (2). To a suspension of 44.5 g (0.93 mol) of washed (hexane) 50% NaH mineral oil dispersion in 50 mL of toluene was added with mechanical stirring under argon a solution of 46.8 g (0.49 mol) of chloroacetic acid in 200 mL of toluene over 1 h. A solution of 100 g (0.41 mol) of 1hexadecanol in 200 mL of toluene was then added over 15 min. The mixture was stirred a t reflux 40 h, cooled to room temperature, and acidified with dilute HCl. The mixture was heated until all solid dissolved. The hot organic layer was dried over MgS04, filtered, and cooled, giving 84.5 g (68%) of 2 as a colorless solid: mp 64-66 "C (lit.'* mp 64.3 "C); NMR (CDCI,) 6 7.78 (b S, 1 H, COpH). 4.18 (s, 2 H, OCHZCOZ), 3.60 (t, 2 H, OCHZCHp), (14) Hato, M.; Shinoda, K.; Miyagoma, T. Bull. Chem. Soc. Jpn. 1976. 49, 1257.
Wissner et al. 1.30 (m, 28 H, (CHz)14),0.93 (m, 3 H, terminal CH,); IR (KBr) 1695 cm-' (C==O);mass spectrum m / z 300 (M'). Anal. (C,,H,O,) C; H: calcd, 12.08; found, 11.64. Z-(Hexadecyloxy)ethanol(3). To a solution of 80 g (0.27 mol) of 2 in 350 mL of T H F was added, dropwise with stirring a t 0 "C under argon, 799 mL of 1 M borane in THF over a 1-h period. After an additional 2 h, 50 mL of acetone was added dropwise to destroy the excess borane followed by 60 mL of HzO. The THF was evaporated, and the residue was heated in a mixture of HzO and CHCl, to dissolve the solids. The organic layer was separated and dried (MgSOJ. The solvent was removed. The residue was recrystallized from CH,OH, giving 70.6 g (93%) of 3 as a colorless solid: mp 42-44 "C; NMR (CDCl,) 6 3.7 (s, 1 H, OH), 3.50 (m, 6 H, OCHz's), 2.18 (m, 2 H, CHzCHzO),1.32 (m, 26 H, (CH2)13), 0.91 (m, 3 H, terminal CH,); IR (KBr) 3275 (OH) cm-'; mass spectrum m / z 255 (M = CHzOH). Anal. (C18H3802)C, H. (Hexadecy1oxy)acetaldehyde (4). A solution of 49.4 g (0.39 mol) of oxalyl chloride in 900 mL of CH2Cl2was cooled to -60 "C, and a solution of 60.8 g (0.78 mol) of MezSO in 175 mL of CHzCIzwas added dropwise with stirring over 0.5 h so that the reaction temperature never exceeded -50 "C. The solution was warmed to -20 "C and maintained a t this temperature as a solution of 44.6 g (0.16 mol) of 3 in 350 mL of CHzClzwas added dropwise over 20 min. The mixture was stirred another 20 min, and 98.2 g of (C2H5),N was added. The mixture was allowed to warm to 25 "C, and HzO was added. The organic layer was separated and washed first with dilute HC1 and then with saturated NaHCO, solution until neutral. The solution was dried (MgS0,-Norite) and filtered through a pad of silica gel. Solvent was removed at reduced pressure at 30 "C, giving 32 g (76%) of 4 as a yellow solid that was not purified further; 4 was stored at low temperature in petroleum ether (30-60 "C): NMR (CDCl,) 6 9.67 (s, 1 H, CHO), 3.99 (9, 2 H, OCHZCHO),3.45 (t, 3 H, CH20), 1.23 (m, 28 H, (CHJ14), 0.88 (m, 3 H, terminal CH3); mass spectrum m / z 420 (M'). l-(Hexadecyloxy)-3-buten-2-ol ( 5 ) . A suspension of 5.67 g (0.23 mol) of Mg was stirred in 25 mL of THF. A few drops of dibromoethane were added. After the evolution of ethylene was observed, another 100 mL of T H F was added, followed by the dropwise addition of a solution of 49.9 g (0.47 mol) of vinyl bromide in 250 mL of THF at a rate that maintained reflux. After all the Mg was consumed, the solution was cooled to 0 "C and a solution of 44.2 g (0.16 mol) of 4 in 125 mL of ether was added dropwise over 0.5 h. After stirring an additional 1 h a t 25 "C, dilute HC1 was added. The T H F was removed, and the residue was extracted with ether. The ether solution was washed with brine and dried (MgS04). The solvent was removed, the residue was chromatographed via HPLC (silica gel, hexane-ether (9:1)), and the product was isolated. Short-path distillation (185-190 "C, 0.2 mm) gave 20.2 g (42%) of 5 as an oil that crystallized on standing: mp 28.5-30 "C; NMR (CDCl,) F 6.15-5.11 (m, 3 H, CH=CHz),4.33 (m, 1H, CH), 3.70-3.15 (m, 4 H, CHzOCHz),2.48 (d, 1H, OH), 1.30 (m, 28 H, (CHz)14),0.91 (m, 3 H, terminal CH3): IR (KBr) 3450 (OH), 1650 (CH=CHz cm-'; mass spectrum m / z 312 (M+). Anal. (C20H4,Oz) C, H. [ [ [ 1-[ (Hexadecyloxy)methyl]-2-propenyl]oxy]methyl]~ benzene (6). To a suspension of 3.35 g (69.9mmol) of NaH (50% mineral oil dispersion) in 75 mL of DMF was added 10.92 g (63.8 mmol) of benzyl bromide and dropwise a solution of 19 g (60.8 mmol) of 5 in 75 mL of DMF. The mixture was stirred overnight and quenched with HzO. The mixture was extracted with ether. The ether solution was dried (MgSO,). Solvent was removed, and the residue was chromatographed via HPLC (silica gel, hexaneCHC1, (8:1)), giving 12 g (49%) of 6 as an oil: NMR (CDCI, 6 7.37 (s, 5 H, C,H5), 6.13-5.18 (m, 3 H, CH=CHz), 4.60 (AB q, 2 H, J = 15 Hz, CHzCBH5),4.05 (m, 1 H, CHI, 3.80-3.28 (m, 4 H, CHzOCHz),1.30 (m, 28 H, (CHz)14),0.91 (m, 3 H, terminal CH,); mass spectrum m / z 311 (M - CHzC6H4).Anal. (Cz7H4,OZ)C, H. 4-(Hexadecy1oxy)-3-(pheny1methoxy)-1 -butanol (7). To a solution of 11.5 g (28.6 mmol) of 6 in 50 mL of THF was added dropwise with stirring under argon 57.1 mL of 1 M borane in THF at 0 "C. After the mixture was stirred 3 h at 25 "C, HzO was slowly added. The mixture was cooled in an ice bath, and 100 mL of 3 N NaOH solution and 100 mL of 30% HzOZwere added with stirring. After 10 min, the mixture was warmed to 25 "C and
Analogues of Platelet Activating Factor stirred 0.5 h. The mixture was poured into ether. The ether solution was washed with brine and saturated NaHS0, solution and dried (MgSO,). Solvent was removed, and the residue was chromatographed via HPLC (silica gel, hexane-ethyl acetate (91)) to give 7.8 g (65%) of 7 as an oil: NMR (CDCl,) 6 7.38 (9, 5 H, CsH5), 4.70 (AB q, 2 H, J = 15 Hz, C H ~ C G H 3.81 ~ ) , (m, 2 H, CH20H), 3.66-3.35 (m, 4 H , CHzOCH2),2.50 (m, 1 H, OH), 2.00-1.11 (m, 28 H, (CH2)14),0.90 (m, 3 H, terminal CH,); IR (neat) 3440 (OH) cm-l; mass spectrum m/z 420 (M+). Anal. (C27H&) H; C: calcd 77.09; found, 78.18. 1-(3-Butenyloxy)hexadecane(8). An aliquot of 73.2 g (1.53 mol) of NaH (50%mineral oil dispersion) was washed with hexane to remove oil, and 1.25 L of DMF was added. The suspension was mechanically stirred under argon as a solution of 100 g (1.39 mol) of 3-buten-1-01 (Aldrich) in 300 mL of DMF was added dropwise over 1.5 h. After the mixture was stirred an additional 1h, 423.5 g (1.39 mol) of 1-bromohexadecanewas added dropwise. The mixture was stirred overnight at 25 "C. HzO was slowly added to destroy excess NaH. The mixture was poured into 1.5 L of H20 and extracted several times with ether. The combined ether solutions were washed with brine and dried (MgS04). Solvent was removed. The residue was distilled first through a short Vigreux column to remove lower boiling side products and then with a Kugelrohr apparatus (l45 "C, 0.05 mm) to give 131.6 g (32%) of 8 as an oil: NMR (CDCl,) 6 6.18-4.87 (m, 3 H, CH= CH2),3.38 (m, 4 H, CH20CH2),2.28 (m, 2 H, allylic CHz), 1.23 (m, 28 H, (CH2)14),0.86 (m, 3 H, terminal CH,); IR (neat) 1635 (CH=CH2) cm-'; mass spectrum m/z 255 (M- C3H5). Anal. (CzoH&) C, H. 1-[ (1-Methyl-2-propenyl)oxy]hexadecane (15). A suspension of 10.57 g (0.22 mol) of NaH (50% mineral oil dispersion, washed with hexane to remove oil) was stirred in 150 mL of DMF under argon as a solution of 16.58 g (0.23 mol) of 3-buten-2-01(13) in 100 mL of DMF was added dropwise over 0.5 h. After gas evolution ceased, 76 g (0.19 mol) of the tosylate of 1-hexadecanol was added, followed by 70 mL of DMF. After stirring overnight, the mixture was poured into water and extracted with petroleum ether. The solution was dried (MgS@,),and solvent was removed. The residue was distilled in a Kugelrohr apparatus (180-185 "C, 0.35 mm) to give 50.9 g (90%) of 15 as a colorless liquid: NMR (CDCl,) 6 6.05-4.98 (m, 3 H, CH=CHz),3.83 (m, 1 H, HCO), 3.40 (m, 2 H, CH20), 1.28 (m, 28 H, (CH2)14), 1.22 (9, 3 H, CH,), 0.91 (m, 3 H, terminal CH,); IR (neat) 1643 (CH=CH2) cm-'; mass spectrum m / z 296 (M'). Anal. (C2oH4,O) C, H. 1-[ (l,l-Dimethyl-2-propenyl)oxy]hexadecane(16). This compound was prepared by a procedure identical with the above using 150 g (0.38 mol) of the tosylate of 1-hexadecanol, 23.6 (0.49 mol) of NaH (50% dispersion), and 55.4 g (0.64 mol) of 14,giving 75.6 g of 16 as a colorless liquid: NMR (CDC1,) 6 5.80 (m, 1 H, CH=CH2), 5.1 (m, 2 H, CH=CHz), 3.25 (t, 2 H, CH20),1.25 (m, 34 H, (CH2)14, C(CH3),), 0.89 (m, 3 H, terminal CH,). Anal. (CziH420) C, €3. [ 1-(Hexadecyloxy)ethyl]oxirane (17). A solution of 46.9 g (0.16 mol) of 15 and 32.2 g (0.21 mol) of m-chloroperbenzoic acid was stirred in 300 mL of CH2C12overnight, and the mixture was filtered. The solvent was removed from the filtrate. The residue was dissolved in a petroleum ether-ether mixture, and the solution was washed with a saturated solution of NaHCO,. Solvent was removed, and the residue was chromatographed via HPLC (silica gel, hexane-ether (9:l)) to give 33 g (67%) of 17 as a colorless oil. TLC (hexane-ether (9:l)) showed two isomers in comparable amounts: NMR (CDC1,) 6 3.85-2.35 (m, 6 H, CH,O's, CHO's), 1.8-1.08 (m, 31 H, (CH2)14,CH,), 0.90 (m, 3 H, terminal CH,). Anal. (C2oH4oOz) C, H. [1-(Hexadecy1oxy)-1-methylethylloxirane(18). This compound was prepared by a method similar to the above from 36.5 g (0.12 mol) of 16 and 23.9 g (0.15 mol) of m-chloroperbenzoic acid, giving 36 g (97%) of 18 which was used in the next step without purification. [2-(Hexadecyloxy)ethyl]oxirane (9). This compound was prepared by a similar method from 100 g (0.34 mol) of 8 and 69 g (0.44 mol) of m-chloroperbenzoic acid, giving 83.4 g (79%) of 9 as an oil: NMR (CDC1,) 6 3.70-3.30 (m, 4 H , CH20CH2),3.05, 2.78 (m, 3 H, epoxide), 2.50 (m, 2 H, CH2CH20),1.30 (m, 28 H, (cHZ)14),0.90 (m, 3 H, terminal CH,); mass spectrum m/z 312 (M'). Anal. (C20H4002) C, H.
Journal of Medicinal Chemistry, 1985, Vol. 28, No. 9 1185
3-(Hexadecyloxy)-1,2-butanediol(19). A solution of 30 g (0.10 "01) of 17 was refluxed in 200 mL of acetic acid containing 0.2 g of p-toluenesulfonic acid (PTSA) for 4.5 h. The solvent was removed at reduced pressure. The residue was stirred in 22 mL of CH,OH, and a solution of 13.44 g (0.34 mol) of NaOH in 14 mL of HzO was added. After stirring for 0.5 h, the solvent was removed and the residue was diluted with H20. The mixture was extracted with ether. The ether solution was washed with brine and dried (MgSO,). Solvent was removed, and the residue was distilled in a Kugelrohr apparatus (200 "C, 0.5 mm) to give 28.9 g (91%) of 19 as a sticky solid: NMR (CDC1,) 6 4.08-3.08 (m, 8 H, OCH2's, OCH's, OH's), 1.73-1.05 (m, 31 H, (CH2)14, CH,), 0.90 (m, 3 H, terminal CH,); IR (KBr) 3300,3425 (OH) cm-'. Anal. (CZOH4203) C, H. 4-(Hexadecyloxy)-1,2-butanediol(lO).This compound was prepared by a method similar to the above using 40 g (0.13 mol) of 9,270 mL of acetic acid, 0.27 g of PTSA, and 17.9 g of NaOH, giving 32 g, (76%) of 10 as a colorless solid: mp 60-61 "C; NMR (CDC1,) 6 4.10-3.18 (m, 9 H, CH20's, CHO, OH's), 2.25 (m, 2 H, CHzCH3),2.05-1.05 (m, 28 H, (CHZll4),0.90 (m, 3 H, terminal CH,); IR (KBr) 3360 (OH) cm-'; mass spectrum m/z 299 (M CH20H). Anal. (C20H4203) C, H. 3-(Hexadecyloxy)-3-methyl-l,2-butanediol (20). A solution of 35 g (0.11 mol) of 18 in 450 mL of MezSO was stirred, and a solution of 30.1 g (0.54 mol) of KOH in 700 mL of HzO was added. The solution was maintained at 110 "C for 8 h. The mixture was cooled, poured into H 2 0 , and extracted with ether. The ether solution was dried (MgS04), and solvent was removed. The residue was chromatographed via HPLC (silica gel, hexane-ethyl acetate (4:1)),giving as the more polar component 18.2 g (49%) of 20 as a colorless solid: mp 30-31 "C; NMR (CDCI,) 6 3.83-3.23 (m, 5 H, CH20's, CHO), 2.73 (b s, 2 H, OH's), 1.88-1.08 (m, 34 H, (CH2)14, (CH3)z),0.90 (m, 3 H, terminal CH,); mass spectrum m / z 283 (M - CH20H);IR (KBr) 3290 (OH) cm-'. Anal. (C2'H4403) C, H. From the earlier fractions was obtained 13 g of a less polar component (CH3(CH2)150C(CH3)2CH(OH)CHz)z0: mp 40-42 "C; mass spectrum m / z 637 (M - HzO, CH,). Anal. (C42H3605) C, H. 3 4Hexadecyloxy )-%(phenylmet hoxy)-1 -butanol (23). A solution of 27.5 g (83.2 mmol) of 19 and 34.8 g (120 mmol) of trityl chloride in 125 mL of dry pyridine was allowed to stand a t room temperature for 48 h. The mixture was poured into HzO and extracted with a mixture (1:l)of ether and petroleum ether (30-60 "C). The organic solution was washed with brine and dried (MgSO,). Solvent is removed, giving 46.6 g (98%) of crude 21 which is used in the next step without additional purification. To a suspension of 5.01 g (100 mmol) of NaH (50% oil dispersion), which was prewashed with hexane, in 200 mL of DMF was added 15.8g (92.3 mmol) of benzyl bromide. The mixture was stirred under argon at 0 "C, and a solution of 46 g (80.3 mmol) of 21 in 50 mL of DMF was added over 0.5 h. The mixture was stirred a t 25 "C overnight, poured into HzO, and extracted with petroleum ether (30-60 "C). The organic layer was dried (MgS04), and solvent was removed. The residue was dissolved in 190 mL of CH30H and 100 mL of THF containing 0.4 g of PTSA. After the mixture was allowed to stand a t 25 "C overnight, the solvent was removed. The residue was dissolved in petroleum ether (30-60 "C); the solution was washed with saturated NaHCO, solution and dried (Na2S04). Solvent was removed, and the residue was chromatographed via HPLC (silica gel, hexaneeethyl acetate (91)) to give 21.8 g (65%) of 23 as an oil. TLC (hexane-ethyl acetate, silica gel) showed two isomers in comparable amounts: NMR (CDC13) 6 7.40 (S,5 H, C&5), 4.71 (m,2 H, CH&&,), 3.98-3.20 (m, 6 H, CHzO's, CHO's), 2.43 (m, 1 H, OH), 2.1C-1.08 (m, 31 H, (CH2)14,CH,), 0.90 (m, 3 H, terminal CH,); IR (neat) 3450 (OH) cm-'. Anal. (C27H4803) C, H. 4-(Hexadecyloxy)-2-(phenylmethoxy)-l-butanol (12). This compound was prepared by a method similar to the above from 30 g (90.8 mmol) of 10 and 37.95 g (130 mmol) of trityl chloride to give 56.7 g of 11 which, on alkylation with 6.17 g (128.5 mmol) of 50% NaH dispersion and 19.5 g (114 mmol) of benzyl bromide, gave, after deprotection, 23 g (61%) of 12 as an oil: NMR (CDCl,) 6 7.35 (5, 5 H, C6H5),4.60 (s, 2 H, CH2C6H5),3.91-3.30 (m, 8 H, CHZO's, CHO, OH), 2.39 (m, 2 H, CHzCH20),2.18-1.11 (m, 28 H, (CH2)14),0.89 (m, 3 H, terminal CH,); IR (neat) 3340 (OH)
1186 cJournal of Medicinal Chemistry, 1985, Vol 28, N o 9 cm-'; mass spectrum miz 391 (M - CH20H). Anal. (C2:H4803) C, H. 3-(Hexadecyloxy)-3-methyl-2(pheny1methoxy)-1 -butanol (24). This compound was prepared in a similar manner to the above from 20 (17.5 g, 50.8 mmol) except that protection of the primary hydroxyl group was accomplished with p-anisylchlorodiphenylmethane (17.9 g, 57.9 mmol) instead of trityl chloride, giving 22 which was alkylated with benzyl bromide (13.5g, 78.9 mmol) and deblocked with Amberlyst 15 ion-exchange resin in methanol-THF, giving 12.1 g (53%) of 24 as an oil: NMR (CDCl,) 6 7.35 (s, 5 H, CsH5), 4.70 (s, 2 H, CH~CGH,), 3.82, 3.45 (m, 5 H, CH20's, CHO), 3.00 (m, 1 H, OH), 1.80-1.18 (m, 28 H, (CHp)14), 0.96 (m, 3 H, terminal CH,). Anal. (C28H5003)C, H. 4-(Hexadecyloxy)-3-(phenylmethoxy)-2-butanol(27). A solution of 9.4 g (73.7 mmol) of oxalyl chloride in 130 mL of CH2C12 was stirred under argon at -70 OC as 11.5 g (147.5 mmol) of MepSO was added dropwise so that the reaction temperature never exceeded -65 "C. A solution of 10 g (24.6 mmol) of 25* in 60 mL of CH2Clzwas added rapidly. The mixture was allowed to warm slowly to -10 OC. The solution was then recooled to -50 "C, and 16.2 g (159.8 mmol) of triethylamine was added. The reaction mixture was warmed to 25 "C and poured into 100 mL of HpO. The organic layer was washed with dilute HCl and a saturated solution of NaHC0,. The organic layer was dried (MgSO,) and solvent was removed, giving 10 g of 26 as an oil that was used without additional purification. To a solution of 26 in 120 mL of dry THF was added, at 5 "C, under argon with stirring, 16.4 mL (49.2 mmol) of 3 M CH3MgBr in ether at a rate such that the temperature never exceeded 8 "C. After 15 min, the mixture was warmed to 25 "C and stirred for 3 h. The solution was cooled to 5 "C, and 50 mL of 1 N HCl was added at a rate that maintained the temperature at 10 "C. The organic phase was separated, and the aqueous phase was extracted with ether. The combined organic solutions were washed with saturated NaHCO, solution and brine and dried (MgSOJ. Solvent was removed, and the residue was chromatographed via HPLC (silica gel, hexane-ether (51)) to give 6.16 g (60%) of 27 as an oil which showed two isomers on TLC in comparable amounts: NMR (CDCl,) 6 7.43 (s, 5 H, C&&, 4.73 (m, 2 H, CH2C6H5), 4.20-3.30 (m, 6 H, CH20's, CHO's), 2.63 (s, 1 H, OH), 1.81-1.06 (m, 31 H, (CHJ14, CHJ, 0.91 (m, 3 H, terminal CH?): IR (neat) 3415 (OH) cm-'. Anal. (C27H4803) C, H. 4-Hydroxy-N,N,N-trimethyl-8-(phenylmethoxy)-3,5,10trioxa-4-phosphahexacosan-1-aminium4-Oxide,Hydroxide, Inner Salt (29). A solution of 7.3 g (17.4 mmol) of 7,7.3g (30.4 mmol) of 28,and 3.07 g (30.4 mmol) of triethylamine in 160 mL of CCl, was stirred for 1.5 h. The mixture was filtered, and solvent was removed. The residue was stirred in a mixture of 300 mL of 0.5 M NaOAc and 300 mL of T H F for 3 h. The T H F was removed a t reduced pressure, and the aqueous solution was acidified with HC1 and extracted with ether. The ether solution was washed with brine and dried (MgS04). The solvent was removed, and the residue was dissolved in a mixture of 100 mL of CH3CN, 90 mL of CHCl? and 50 g of anhydrous (CH&N. The solution was refluxed €or 20 h. The solvent was removed, and the residue was stirred in 100 mL CH30H containing 2.7 g of Ag2C03for 2 h. The mixture was filtered, and the solvent was removed. The residue was chromatographed on silica gel, eluting first with CHC13-CHJOH (7:3) to remove the more mobile impurities and then with CHC1,-CH30H-Hz0 (70:30:5) to elute product that was triturated with ether to give 6.31 g (62%) of 29 as a white powder with no well-defined melting point: NMR (CDC1,-CD30H) 6 7.30 (m, 5 H, C6HS),4.60 (AB q, 2 H. J = 15Hz, CH2C&), 4.38-3.21 (7, 11 H, a to 0 and N), 3.10 (s, 9 H, N(CH,),), 1.85 (m, 2 H, CH2CH20),1.23 (m, 28 H, (CH2)14),0.84 (m, 3 H, terminal CH,); mass spectrum (FAB) m / r 586 (M + H). Anal. (C,2H6006PN.H20)c, H, N, P. 4-Hydroxy-N,N,N-trimethyl-7-(phenylmethoxy)-3,5,10trioxa-4-phosphahexacosan1-aminium 4-Oxide,Hydroxide, Inner Salt (30). This compound was prepared by a method similar to the above from 10.0 g (23.8 mmol) of 12 to give 9.33 g (67%) of 30 as a white powder with no well-defined melting point: NMR (CDCl,-CD,OD) 6 7.35 (m, 5 H , C&), 4.62 (AB q, 2 H, CH2C6H5),4.17 (m, 2 H, POCH2), 4.03, 3.89 (m, 2 H, CH20P),3.58 (m, 1 H, CH), 3.56-3.29 (m, 6 H, CH20CH2,CH2N), 3.09 (s,'3 i t i N(CHJ,). 1 83 (111. 2 14 CHCH,CH,). 1.53 im. 2 H.
Wissner et al. OCH,CHJ, 1.25 (m, 26 H, (CH2)13),0.88 (m, 3 H, terminal CH,); mass spectrum (FAB) m / z 586 (M + H). Anal. (Cs2Hso06NP*HZO)C, H, P, N. 4-Hydroxy-N,N,N,8-tetramethyl-7-(phenylmethoxy)3,5,9-trioxa-4-phosphapentacosan-l-aminium 4-Oxide,H y droxide, Inner Salt (31). This compound was prepared by a method similar to the above from 20 g (47.5 mmol) of 23 to give 10.4 g (39%) of 31 as a colorless foam: NMR (CDC1,-CD,OD) 8 7.28 (m, 5 H, CsH5), 4.70 (m, 2 H, CHZC~H,), 4.00-3.30 (m, 10 H, a to 0 and N), 3.15 (s, 9 H, N(CH,),), 1.64 (m, 2 H, CH2CH20), 1.24 (m, 29 H, (CH2)13,CH,), 0.89 (m, 3 H, terminal CH,); mass spectrum (FAB) m / z 586 (M + H). Anal. (C32H600BPN.4H20) C, N, P; H: calcd, 10.42; found, 9.10. 4-Hydroxy-N,N,N,8,8-pentamethyl-'l-(phenylmethoxy)3,5,9-trioxa-4-phosphapentacosan-l-aminium 4-Oxide,H y droxide, Inner Salt (32). This compound was prepared by a similar method from 11 g (25.3 mmol) of 24 to give 5.4 g (36%) of 32 as a white powder with no well-defined melting point: NMR (CDCl,-CD,OD) 6 7.37 (m, 5 H, C6Hj),4.80 (AB q, 2 H, CH2C6H6), 4.16 (m, 2 H, POCH2),4.25,3.92 (m, 2 H, CH20P), 3.60 (m, 1 H, CHO), 1.25 (m, 26 H, (CH2)13), 1.25 (s, 3 H, CH,), 0.88 (m, 3 H, terminal CH,); mass spectrum (FAB) m / z 600 (M + H). Anal. (C3&&j&jPN.'/2H20) C, H, N. 4-Hydroxy-N,N,N,6-tetramethyl-7-(phenylmethoxy)3,5,9-trioxa-4-phosphapentacosan-l-aminium 4-Oxide,H y droxide, Inner Salt (33). This compound was prepared by a similar method from 4.8 g (11.4 mmol) of 27 except that the reaction with 28 was allowed to proceed for 48 h, giving 3.6 (54%) of 33 as a colorless powder with no well-defined melting point: NMR (CDC1,-CD,OD) 13 7.20-7.52 (m, 5 H, C&5), 4.78-4.60 (AB q, 2 H, CH2C&j), 4.54-4.12 (m, 3 H, CHOP03CHz),4.00-3.36 (m, 7 H, CH2N, CHO, CHz0CH2),3.12 (s, 9 H, N(CH3),), 1.59 (m, 2 H, CH2CH20),1.26 (m, 29 H, (CHJ13, CH3), 0.88 (m, 3 H, terminal CH,); mass spectrum (FAB) m / z 586 (M + H). Anal. ( C , ~ H , O O ~ P N . ~ HC, ~O H,) N, P. 4,8-Dihydroxy-NJVJVN-trimethyl-3,5,10-trioxa-4-phosphahexacosan-1-aminium4-Oxide,Hydroxide, Inner Salt (34). A solution of 5.6 g (9.6 mmol) of 29 in 35 mL of CH3C02Hand 35 mL of CH,OH containing 0.5 g of 5% Pd/C catalyst was subjected to hydrogenolysis in a Parr shaker for 17 h. The catalyst was removed by filtration, and the solvent was removed, giving 4.7 g (99%) of 34 that was used in the next step without additional purification. 4,7-Dihydroxy-NJVJVN-trimethyl-3,5,lO-trioxa-4-phosphahexacosan-1-aminium4-Oxide,Hydroxide, Inner Salt (35). This compound was prepared from 8.8 g (15 mmol) of 30 and 0.9 g of 5% Pd/C by a similar method to give 7.5 g (100%) of 35 as a white powder after trituration with ether: NMR (CDC13-CD,OD) 6 4.30 (m, 2 H, POCH,), 4.08-3.71 (m, 3 H, CH20P, CHI, 3.66 (m, 2 H, CH2N),3.6C-3.32 ( m , 4 H, CH20CH2),3.25(s,9 H, N(CH,),), 1.71 (m, 2 H, CHCH2CH2),1.53 (m, 2 H, OCH2CH2), 1.25 (m, 26 H, (CH2)13), 0.88 (m, 3 H, terminal CH,); IR (KBr) 3210 (OH) cm-'; mass spectrum (FAB) m / z 496 (M + H). Anal. (C2jH~$K"gl1/2H2O) C, H, N, P. 4,7-Dihydroxy-N,N,N,%tetramethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium4-Oxide,Hydroxide, Inner Salt (36). This compound was prepared by a similar method from 10 g (17.1 mmol) of 31 and 1.0 g of 5% Pd/C to give 7.4 g (88%) of 36 as a thick oil: mass spectrum (FAB) m / z 496 (M + H); IR (neat) 3230 (OH) cm-'. 4,7-Dihydroxy-N,N,N,8,8-pentamethyl-3,5,9-trioxa-4phosphapentacosan-1-aminium 4-Oxide,Hydroxide, Inner Salt (37). This compound was prepared by a similar method from 5 g (8.3 mmol) of 32 and 0.5 g of 5% Pd/C to give after precipitation with ether 3.93 g (92%) of 37 as a white powder with no well-defined melting point: NMR (CDCl,-CD,OH) 6 4.28 (m, 2 H, POCH2), 4.13, 3.82 (m, 2 H, CH20P), 3.64 (m, 3 H, CHO, CH2N), 3.36 (t, 2 H, OCH2), 3.06 (9, 9 H, N(CH,),), 1.49 (m, 2 H, CH2CHz0),1.24 (m, 26 H, (CH2)13),1.23 (s, 3 H, CH,), 1.16 (s, 3 H, CH,), 0.88 (m, 3 H, terminal CH,); mass spectrum (FAB) m / z 552 (M H). Anal. (CZ8Hj8O7PN-H20)C, H, P, N. 4,7-Dihydroxy-N,N,N,6-tetramet hyl-3,5,9-trioxa-4-p hosphapentacosan-1-aminium4-Oxide,Hydroxide, Inner Salt (38). This compound was prepared by a similar method from 3.5 g (5.97 mmol) of 33 and 0.6 g of 5% Pd/C, giving after precipitation with ether 2.69 g (91%) of 38 as a white powder with
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Journal of Medicinal Chemistry, 1985, Vol. 28, No. 9 1187
Analogues of Platelet Activating Factor
no well-defined melting point: IR (KBr) 3420 (OH) cm-'; mass anhydride, and 5.96 g (58.9 mmol) of triethylamine except that spectrum (FAB) m/z 496 (M + H). Anal. (C2&I&6PN*1/2H20) the reaction mixture was refluxed for 15 h, giving, after precipitation with ether, 1.0 g (31%) of 42 as a colorless powder with C, H, N, P. 8-(Acetyloxy)-4-hydroxy-~,N,N-trimethyl-3,5,lO-trioxa-no definite melting point: NMR (CDC1,-CD,OD) 6 5.12 (m, 1 4-phosphahexacosan-1-aminium 4-Oxide, Hydroxide, I n n e r H, CHO), 4.24,3.97 (m, 4 H, CHZ0PO3CH2), 3.59 (m, 2 H, CH2N), Salt (39). A solution of 4.5 g (9.1 mmol) of 34, 23.2 g (230 mmol) 3.57 (s, 9 H, N(CH,),), 3.24 (t, 2 H, OCHZ), 3.21 (s, 3 H, COCH,), of acetic anhydride, and 9.2 g (90.8mmol) of triethylamine in 250 1.46 (m, 2 H, OCHzCH2),1.26 (m, 29 H, (CH2)13,CH,), 1.17 (s, mL of CHC1, was refluxed for 4 h. Solvent and excess anhydride 3 H, CH,), 0.88 (m, 3 H, terminal CH,); IR (KBr) 1735 (C=O) were removed a t reduced pressure. The residue was chromatocm-'; mass spectrum (FAB) m / z 552 (M + H). Anal. (C28H,graphed on silica gel (250 mL dry volume), eluting first with O;.PN.H,O) C, H, N, P. CHCl,-CH,OH (7:3) to remove more mobile impurities and then 7-(Acetyloxy)-4-hydroxy-N,N,N,6-tetramethy1-3,5,9-triwith CHC13-CH30H-Hz0 (70:305) to elute product. The product oxa-4-phosphapentacosan1-aminium 4-Oxide, Hydroxide, was precipitated with ether, giving 3.6 g (74%) of 39 as a white I n n e r salt (43). This compound was prepared by a similar powder with no well-defined melting point: NMR (CDC13-Cmethod from 2 g (4 mmol) of 38, 10.3 g (101 mmol) pf acetic D,OD) 6 5.16 (m, 1 H, CHO), 4.25 (m, 2 H, POCH2), 3.93 (m, 2 anhydride, and 4.08 g (40.3 mmol) of triethylamine to give, after H, CHzOP), 3.63 (m, 2 H, CHzN), 3.50 (m, 4 H, CHz0CH2),3.24 precipitation with ether, 1.13 g of (52%) of 43, a mixture of (s,9 H, N(CH,),), 2.07 (s, 3 H, COCH3),1.92(m, 2 H, CHCHzCH2), isomers, as a colorless powder with no definite melting point: 1.54 (m, 2 H, CHzCH20),1.26 (m, 26 H, (CHz)13),0.88 (m, 3 H, NMR (CDC1,-CD,OD) 6 5.17, 5.03 (m, 1 H, CHOAc), 4.44, 4.24 terminal CH,); IR (KBr) 1730 (C=O) cm-'; mass spectrum (FAB) (m, 3 H, CHOP03CHz),3.72-3.33 (m, 6 H, CH2N, CH20CH2), m/z 538 (M + H). Anal. (C27H5607PN.'/2H20) C, H, N, P. 3.22, 3.21 (s, 9 H, N(CH3),), 2.09, 2.08 (s, 3 H, COCH,), 1.52 (m, 7-(Acetyloxy)-4-hydroxy-N,N,N-trimethyl-3,5,lO-trioxa- 2 H, CH2CH20),1.23 (m, 29 H, (CH,),,, CH,), 0.88 (m, 3 H, 4-phosphahexacosan-1-aminium 4-Oxide, Hydroxide, I n n e r terminal CH,); IR (KBr) 1738 (C=O) cm-'; mass spectrum (FAB) Salt (40). This compound was prepared by a similar method from m/z 538 (M + H). Anal. (C27H5607PN.21/zHz0) C, H, N, P. 5.0 g (10.0 mmol) of 35, 25 g (245 mmol) of acetic anhydride, and 10.2 g (100 mmol) of triethylamine, giving 2.88 g (53%) of 40 as Acknowledgment. We thank Dr. L. Gehrlein and staff a colorless powder with no well-defined melting point: NMR for microanalytical determinations, Dr. M. Siege1 and G. (CDCl,-CD,OD) 6 5.14 (m, 1 H, CH), 4.28 (m, 2 H, POCHz), 3.93 (m, 2 H, CH20P),3.65 (m, 2 H, CH2N),3.40 (m, 4 H, CH20CH2), 0. Morton and staff for spectral data, R. A. Gabel for his 3.23 (s, 9 H, N(CH,),), 2.05 (s, 3 H, COCH,), 1.87 (m, 2 H, assistance with the hypotension assay, and D. R. Nytko CHCH2CHz),1.52 (m, 2 H, OCH2CHz),1.26 (m, 26 H, (CH2)13), for her assistance with the platelet aggregation assay. 0.88 (m, 3 H, terminal CH,); IR (KBr) 1735 (C=O) cm-'; mass spectrum (FAB) m/z 538 (M + H). Anal. (C27H5607NP.HzO) C, H, N; P: calcd, 5.57; found, 6.20. Registry No. (&)-la,97277-41-3;2, 58210-02-9;3, 2136-71-2; 7 4Acetyloxy)-4-hydroxy-N,N,N,8-tetramethyl-3,5,9-tri- 4, 60956-71-0; (&)-5, 97150-02-2; (*)-6, 97150-03-3; (*)-7, oxa-4-phosphapentacosan-1-aminium a-Oxide, Hydroxide, 97232-51-4;8,97150-04-4; (&)-9,97150-05-5; (*)-lo, 97150-06-6; Inner Salt (41). This compound was prepared from 6.4 g (12.9 (&)-11,97170-24-6; (*)-12,97150-07-7; 13,598-32-3; 14, 115-18-4; mmol) of 36, 32.95 g (320 mmol) of acetic anhydride, and 13.1 15, 97150-08-8; (*)-16, 97150-09-9; 17, 97150-10-2; (*)-18, g (130 mmol) of triethylamine, giving, after precipitation with 97150-11-3; 19, 97150-12-4; (*)-20, 97150-13-5; 21, 97150-14-6; ether, 4.6 g (66%) of 41, a mixture of two isomers in equal (*)-22,97170-25-7; (&)-23(isomer l), 97150-15-7; (*)-23 (isomer amounts, as a white powder with no well-defined melting point: 2), 97150-16-8; (&)-24,97150-17-9; (*)-25, 18678-94-9; (*)-26, NMR (CDC1,-CD,OD) 6 5.12, 5.02 (m, 1 H, CHOAc), 4.33-3.35 97232-52-5; (&)-27 (isomer l),97150-18-0; (&)-27 (isomer 2), (m, 9 H, a to 0 and N), 3.34 (s, 9 H, N(CH,),), 2.09, 2.08 (s, 3 97150-19-1;28, 4167-02-6; (&)-29,97150-20-4;(*)-30,97150-21-5; H, CH,CO), 1.49 (m, 2 H, CH2CH20),1.06 (m, 28 H , (CHZ)'J, 31, 97150-22-6; (&)-32, 97150-23-7; 33, 97150-24-8; (*)-34, 1.14, 1.12 (d, 3 H, CH,), 0.88 (m, 3 H, terminal CH,); IR (KBr) 97150-25-9; (&)-35,97150-26-0;36,97150-27-1; (*)-37,97150-28-2; 1725 (C=O) cm-'; mass spectrum (FAB) m/z 538 (M H). Anal. 38, 97150-29-3; (&)-39, 97150-30-6; (*)-40, 97150-31-7; (*)-41 (isomer l ) , 97150-32-8; (&)-41 (isomer 2), 97150-33-9; (*)-42, (Cz,H&$'N) C, H, N, P. 7-(Acetyloxy)-4-hydroxy-N,N,N,8,8-pentamethyl-3,5,9- 97150-34-0; (&)-43 (isomer l ) , 97232-53-6; (*)-43 (isomer 2), trioxa-4-phosphapentacosan-1-aminium 4-Oxide, Hydroxide, 97232-54-7;ClCHVCO,H, 79-11-8: CH,=CH(CH,),OH. 627-27-0: I n n e r salt (42). This compound was prepared by a similar CH3(CH2)liBr,lf2-Si-3;' CH3(CH2)156S02CsH,M&, 6068-28-61 method from 3 g (5.9 mmol) of 37, 15 g (150 mmol) of acetic CH,(CH,)i,OH, 36653-82-4.
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